Probing teleost eye development by lens transplantation.

摘要

Experimental manipulation and other lines of evidence indicate that the lens plays a prominent role in the growth and differentiation of the vertebrate eye. Here we describe a lens transplantation method for studying the role of the lens in teleost eye development. The method involves three steps: (1) preparing embryos for the operations by embedding them in agar, (2) microsurgery with tungsten needles to remove the lens from a donor embryo and insert it into the optic cup of a host embryo lacking its own lens, and (3) a recovery period allowing surface ectoderm to close over the wound left by insertion of the lens into the host embryo. A movie illustrating the method can be found at http://www.life.umd.edu/labs/jeffery. A troubleshooting guide and summary of assays for evaluating the development of the transplanted lens and its effects on other eye parts, including the retina, are presented. Finally, some current applications of the lens transplantation method are briefly described: (1) determination of the autonomy of zebrafish lens mutants and (2) investigation of the role of the lens in eye degeneration in the cavefish Astyanax. The transplantation method will help characterize the mechanisms through which vertebrate eye development is regulated by the lens.